Abstract：
:We have examined in detail the kinetics of binding of the serpin alpha 2-antiplasmin to the serine proteases alpha-chymotrypsin and plasmin. These represent model systems for serpin binding. We find, in contrast to earlier published results with alpha 2-antiplasmin and plasmin, that binding is reversible, and slow binding kinetics can be observed, under appropriate conditions. Binding follows a two-step process with both enzymes, with the formation of an initial loose complex which then proceeds to a tightly bound complex. In the absence of lysine and analogues, equilibrium between alpha 2-antiplasmin and plasmin is achieved rapidly, with an overall inhibition constant (Ki') of 0.3 pM. In the presence of tranexamic acid or 6-aminohexanoic acid, lysine analogues that mimic the effects of fibrin, plasmin binding kinetics are changed such that equilibrium is reached slowly following a lag phase after mixing of enzyme and inhibitor. The Ki' is also affected, rising to 2 pM in the presence of 6-aminohexanoic acid concentrations above 15 mM. Thus extrapolation to the in vivo situation indicates that complex formation in the presence of fibrin will be delayed, allowing a burst of enzyme activity following plasmin generation, but a tight, pseudoirreversible complex will result eventually. Chymotrypsin is more weakly inhibited by alpha 2-antiplasmin, exhibiting an overall Ki' of 0.1 nM, after two-stage complex formation. The inhibition constant for the initial loose complex (Ki) is very similar for both enzymes. The difference in binding strength between the two enzymes is accounted for by the dissociation rate constant of the second step of complex formation.(ABSTRACT TRUNCATED AT 250 WORDS)
journal_name
Biochemistryjournal_title
Biochemistryauthors
Longstaff C,Gaffney PJdoi
10.1021/bi00218a014subject
Has Abstractpub_date
1991-01-29 00:00:00pages
979-86issue
4eissn
0006-2960issn
1520-4995journal_volume
30pub_type
杂志文章相关文献
BIOCHEMISTRY文献大全abstract：:Ethanol, in vitro, is known to perturb the molecular order of the phospholipids in biological membranes, while chronic ethanol exposure, in vivo, leads to resistance to disordering. Such changes have usually been measured by electron spin resonance, utilizing fatty acid spin probes. The use of such probes is controver...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00346a012
更新日期：1985-12-03 00:00:00
abstract：:Among hormone-inducible transcription factors, estrogen receptors (ERs) play important roles in tissue growth and differentiation, via either direct or indirect binding, in the nucleus, to specific DNA targets called estrogen responsive elements (EREs), or through nongenomic pathways. In humans, two estrogen receptor ...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/acs.biochem.5b00026
更新日期：2015-06-30 00:00:00
abstract：:IFABP is a small beta-barrel protein with a short helix-turn-helix motif near the N-terminus that is thought to participate in the regulation of the uptake and delivery of fatty acids. In a previous work, we detected by near UV circular dichroism a reversible conformational transition of this protein occurring between...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi020680d
更新日期：2003-06-24 00:00:00
abstract：:The binding of various forms of vanadate to myosin and myosin subfragment 1 (S-1) was studied by 51V NMR at increasing vanadate concentrations between 0.06 and 1.0 mM. The distribution of the various forms of vanadate in the solution depended on the total concentration of vanadate. At low concentrations, the predomina...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00490a029
更新日期：1990-09-25 00:00:00
abstract：:The dependence of the activity of recombinant activated human protein C (r-APC) on each of its nine gamma-carboxyglutamic (Gla) residues (sequence positions 6, 7, 14, 16, 19, 20, 25, 26, and 29) has been assessed in purified systems and in plasma using r-mutants in which each Gla residue of r-APC was individually alte...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00173a033
更新日期：1994-02-22 00:00:00
abstract：:Bacterial NO synthase (NOS)-like proteins such as that from Bacillus subtilis (bsNOS) share a high degree of structural homology with the oxygenase domain of mammalian NOSs (mNOSs), but biochemical studies have yet failed to establish that they are specifically capable of producing NO. To better understand the actual ...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi051710q
更新日期：2006-02-07 00:00:00
abstract：:A conformational transition is described for the polypeptide, gramicidin A, in which a dimer that forms a left-handed intertwined antiparallel helix is converted to a single-stranded amino terminus to amino terminus right-handed helix. The starting structure is determined here by solution NMR methods while reference i...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00152a019
更新日期：1992-09-22 00:00:00
abstract：:In order to gain insight into the structure of human parathyroid hormone (hPTH), four fragments [hPTH(1-34), hPTH(18-48), hPTH(28-48), and hPTH(53-84)], which cover all regions of the intact hormone, have been investigated by CD and NMR spectroscopy in combination with distance geometry, and restrained molecular dynam...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00173a010
更新日期：1994-02-22 00:00:00
abstract：:The slowly hydrolyzed ATP analog adenosine 5'-(gamma-thiotriphosphate) (ATP gamma S) has been used in many studies of the muscle motor protein myosin in order to form a stable "weak binding" state analogous to the actin-S1-ATP complex However, the results from studies using ATP gamma S do not always agree with the res...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00049a018
更新日期：1995-12-12 00:00:00
abstract：:The 70-kDa heat-shock protein (Hsp70) has been cloned and sequenced from bovine cardiac muscle. On the basis of sequence features, the gene corresponds to the cytoplasmic form of Hsp70. This cardiac Hsp70 cDNA clone has an open reading frame of 1926 bp coding for 641 amino acids and a predicted molecular mass of 70.25...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi049036k
更新日期：2004-10-26 00:00:00
abstract：:The fosfomycin (1) resistance proteins FosA and FosX in pathogenic microorganisms are related to a catalytically promiscuous progenitor encoded in a phn operon in Mesorhizobium loti. The mlr3345 gene product (FosX(Ml)) from M. loti has a very low epoxide hydrolase activity and even lower glutathione transferase activi...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi900078q
更新日期：2009-03-10 00:00:00
abstract：:The intense intrinsic fluorescence emissions from several clinically relevant camptothecin drugs have been exploited in order to determine (1) the structural basis of drug binding to lipid bilayers, (2) the lipid bilayer stability of each drug's alpha-hydroxylactone moiety, a pharmacophore which is essential for antit...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00071a010
更新日期：1993-05-25 00:00:00
abstract：:Kinetic studies of reactions between fluorescein and antifluorescein antibody produced during early, intermediate, and late stages of the immune response have been carried out utilizing both fluorescence intensity and polarization measurements in the static (time constant similar to 5 sec) and in the stopped-flow mode...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00688a009
更新日期：1975-08-26 00:00:00
abstract：:1H NMR spectroscopy of the isotropically shifted signals in cobalt carboxypeptidase, CoCPD, permits a direct and selective detection of protons belonging to the residues liganded to the metal. The chemical shift of these protons in the free enzyme and enzyme-inhibitor complexes with changing pH monitors the state of i...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00130a015
更新日期：1992-04-21 00:00:00
abstract：:The effects of various compounds such as the transport substrate ADP and the transport inhibitors carboxyatractyloside (CATR) and bongkrekic acid (BKA) on the labeling of cysteine residues in the ADP/ATP carrier of bovine heart submitochondrial particles by the SH reagent eosin-5-maleimide (EMA) were studied. Of the f...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00198a019
更新日期：1994-08-16 00:00:00
abstract：:CD6 is a member of the scavenger receptor cysteine rich protein superfamily (SRCRSF). This family includes many cell surface proteins whose three-dimensional structures and functions are presently not well understood. The extracellular region of CD6 includes 3 SRCR domains. The membrane proximal SRCR domain specifical...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi962560+
更新日期：1997-03-04 00:00:00
abstract：:The ionization and phase behavior of 1,2-dipalmitoyl-sn-glycero-3-phosphoserine have been investigated under a variety of condtions by several different methods. As measured by turbidity changes, the temperature of the crystal-liquid crystal phase transition of this lipid is influenced by pH and mono- and divalent ca...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00649a025
更新日期：1976-02-24 00:00:00
abstract：:The metalloregulatory protein MntR from Bacillus subtilis acts as a transcriptional regulator of manganese homeostasis. MntR is a member of a subfamily of DtxR-related proteins that perform analogous regulatory functions in a variety of pathogenic organisms. Metal ions activate MntR to bind DNA and repress the transcr...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi0350248
更新日期：2003-11-04 00:00:00
abstract：:Guanidinium chloride- (GdmCl-) induced unfolding of beta-lactamase has been investigated by a combination of size-exclusion chromatography (SEC-FPLC) and usual optical methods. It has been shown that at low temperatures this protein unfolds through two equilibrium intermediates. The first of these intermediates is the...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00176a006
更新日期：1994-03-15 00:00:00
abstract：:Phytate is an antinutritional factor that influences the bioavailability of essential minerals by forming complexes with them and converting them into insoluble salts. To further our understanding of the chemistry of phytate's binding interactions with biologically important metal cations, we determined the stoichiome...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi1010249
更新日期：2010-11-30 00:00:00
abstract：:Streptomyces avermitilis, an industrial organism responsible for the production of the anthelminthic avermectins, harbors a 13.4 kb gene cluster containing 13 unidirectionally transcribed open reading frames corresponding to the apparent biosynthetic operon for the sesquiterpene antibiotic pentalenolactone. The advanc...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi060419n
更新日期：2006-05-16 00:00:00
abstract：:Reactivation of tetrameric porcine skeletal muscle lactic dehydrogenase after dissociation and extensive unfolding of the monomers by 6 M guanidine hydrochloride (Gdn . HCl) is characterized by sigmoidal kinetics, indicating a complex mechanism involving rate-limiting folding and association steps. For analysis of the...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00260a007
更新日期：1982-08-17 00:00:00
abstract：:A panel of 10 hybridomas was assembled to assess the influence of various genetic and biological factors upon glycosylation of secreted monoclonal IgG. After exhaustive Pronase digestion of IgG, glycopeptides were characterized chromatographically by apparent size, charge, and concanavalin A (Con A)-Sepharose and Lens...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00429a065
更新日期：1989-02-07 00:00:00
abstract：:The EcDos protein belongs to a group of heme-based sensors that detect their ligands with a heme-binding PAS domain. Among these various heme-PAS proteins, EcDos is unique in having its heme iron coordinated at both axial positions to residues of the protein. To achieve its high affinities for ligands, one of the axia...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi025845x
更新日期：2002-07-02 00:00:00
abstract：:Three crystal structures have been determined of active site specific substituted Cd(II) horse liver alcohol dehydrogenase and its complexes. Intensities were collected for the free, orthorhombic enzyme to 2.4-A resolution and for a triclinic binary complex with NADH to 2.7-A resolution. A ternary complex was crystall...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00346a070
更新日期：1985-12-03 00:00:00
abstract：:A striking feature of the CLIC (chloride intracellular channel) protein family is the ability of its members to convert between a soluble state and an integral membrane channel form. Direct evidence of the structural transition required for the CLIC protein to autonomously insert into the membrane is lacking, largely ...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi100111c
更新日期：2010-06-29 00:00:00
abstract：:A series of monoclonal antibodies specific for distinguishable epitopes in chromosomal protein histone H5 were obtained from mice immunized with either free H5 or H5 . RNA complexes. The antibodies elicited by H5 could be distinguished from those elicited by H5 . RNA by their binding to native or acid-denatured H5, by...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00310a012
更新日期：1984-07-17 00:00:00
abstract：:Understanding the process of amyloidogenesis is important for the future treatment of misfolding-based diseases, such as Alzheimer's, spongiform encephalopathies, and other important disorders affecting humans. In this work, we have used one of the best-characterized models for folding and misfolding, the activation d...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi900960s
更新日期：2009-11-10 00:00:00
abstract：:Imidazolonepropionase (HutI) (imidazolone-5-propanote hydrolase, EC 3.5.2.7) is a member of the amidohydrolase superfamily and catalyzes the conversion of imidazolone-5-propanoate to N-formimino-L-glutamate in the histidine degradation pathway. We have determined the three-dimensional crystal structures of HutI from A...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi800180g
更新日期：2008-05-20 00:00:00
abstract：:High-resolution proton nuclear magnetic resonance (1H NMR) has been used to study the conformation and dynamics of biosynthetic analogues of actinomycin D that have been substituted at one or both of the 3'-amino acids with azetidine-2-carboxylic acid, pipecolic acid, or 4-ketoproline for one or both of the prolines. ...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00540a007
更新日期：1982-05-25 00:00:00
